Quantitative profiling of inflammatory and pro-resolving lipid mediators in human adolescents and mouse plasma using UHPLC-MS/MS
Author:
Hartling Ivan12ORCID, Cremonesi Alessio1, Osuna Ester3, Lou Phing-How4, Lucchinetti Eliana5, Zaugg Michael45, Hersberger Martin12ORCID
Affiliation:
1. Division of Clinical Chemistry and Biochemistry , Children’s Research Center, University Children’s Hospital Zurich, University of Zurich , Zurich , Switzerland 2. Zurich Centre for Integrative Human Physiology , University of Zurich , Zurich , Switzerland 3. Human Nutrition Laboratory, Department of Health Sciences and Technology , ETH Zurich , Zurich , Switzerland 4. Department of Anesthesiology and Pain Medicine , Cardiovascular Research Centre, University of Alberta , Edmonton , Canada 5. Department of Pharmacology , University of Alberta , Edmonton , Canada
Abstract
Abstract
Objectives
Lipid mediators are bioactive lipids which help regulate inflammation. We aimed to develop an ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method to quantify 58 pro-inflammatory and pro-resolving lipid mediators in plasma, determine preliminary reference ranges for adolescents, and investigate how total parenteral nutrition (TPN) containing omega-3 polyunsaturated fatty acid (n-3 PUFA) or n-6 PUFA based lipid emulsions influence lipid mediator concentrations in plasma.
Methods
Lipid mediators were extracted from plasma using SPE and measured using UHPLC-MS/MS. EDTA plasma was collected from healthy adolescents between 13 and 17 years of age to determine preliminary reference ranges and from mice given intravenous TPN for seven days containing either an n-3 PUFA or n-6 PUFA based lipid emulsion.
Results
We successfully quantified 43 lipid mediators in human plasma with good precision and recovery including several leukotrienes, prostaglandins, resolvins, protectins, maresins, and lipoxins. We found that the addition of methanol to human plasma after blood separation reduces post blood draw increases in 12-hydroxyeicosatetraenoic acid (12-HETE), 12-hydroxyeicosapentaenoic acid (12-HEPE), 12S-hydroxyeicosatrienoic acid (12S-HETrE), 14-hydroxydocosahexaenoic acid (14-HDHA) and thromboxane B2 (TXB2). Compared to the n-6 PUFA based TPN, the n-3 PUFA based TPN increased specialized pro-resolving mediators such as maresin 1 (MaR1), MaR2, protectin D1 (PD1), PDX, and resolvin D5 (RvD5), and decreased inflammatory lipid mediators such as leukotriene B4 (LTB4) and prostaglandin D2 (PGD2).
Conclusions
Our method provides an accurate and sensitive quantification of 58 lipid mediators from plasma samples, which we used to establish a preliminary reference range for lipid mediators in plasma samples of adolescents; and to show that n-3 PUFA, compared to n-6 PUFA rich TPN, leads to a less inflammatory lipid mediator profile in mice.
Publisher
Walter de Gruyter GmbH
Subject
Biochemistry, medical,Clinical Biochemistry,General Medicine
Reference53 articles.
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